CN112616556A - Method for improving phellinus igniarius cultivation efficiency through laboratory intelligent cultivation management - Google Patents
Method for improving phellinus igniarius cultivation efficiency through laboratory intelligent cultivation management Download PDFInfo
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Abstract
The invention discloses a method for improving the cultivation efficiency of phellinus igniarius through intelligent cultivation management in a laboratory, which comprises the following steps: quality improvement and optimization of a phellinus igniarius mother strain solid plate culture medium, intermittent propagation culture of phellinus igniarius liquid strains, optimization of a culture medium of mulberry twig bag materials, intelligent fungus growing, fruiting cultivation management and the like. The invention has short culture period of phellinus igniarius mycelium and sporocarp, high biotransformation rate and high content of polysaccharide, triterpene and other bioactive components. The invention remotely controls the intelligent management of a laboratory through a mobile phone, remotely, flexibly and accurately controls the cultivation conditions all day long, and shortens the bag filling time of hyphae to about 40 d; the culture period of the phellinus igniarius sporocarp after the fungus bag is opened is shortened to about 70 days, and compared with the culture method in the prior art, the culture method is shortened by more than 60 days.
Description
Technical Field
The invention relates to the technical field of medicinal edible fungus cultivation, in particular to a method for improving the cultivation efficiency of phellinus igniarius through intelligent cultivation management in a laboratory.
Background
Phellinus linteus is a rare traditional Chinese medicinal material growing in the mulberry, has been recorded in detail in Ben Cao gang mu, Chinese medicine dictionary and Shen nong Ben Cao Jing as a traditional Chinese medicine for over 2000 years, and is described as 'take a light body for a long time without aging and prolong life' in Shen nong Ben Cao Jing, so that Phellinus linteus has a name of 'forest gold'. At present, many organizations in China keep phellinus igniarius strains and carry out series research. The anticancer effect of phellinus igniarius was first published by the japan national cancer research center in 1968, and the experiment shows that the water extract of the wild phellinus igniarius sporophore has an inhibitory effect on mouse sarcoma S180 tumor cells, the proliferation inhibition rate is as high as 96.7%, and the phellinus igniarius sporophore is the first in dozens of bacteria. The effective components of Phellinus Linteus mainly contain polysaccharide, flavone and triterpene, and have effects of treating prostatitis, treating gout, enhancing immunity, resisting oxidation, etc. In recent years, medical research shows that phellinus igniarius polysaccharides have the functions of obviously inhibiting tumor growth and metastasis, have no toxic or side effect on human bodies, and are the fungi with the highest efficiency in the field of biological cancer treatment internationally acknowledged at present. In recent years, large pharmaceutical factories at home and abroad invest huge capital for developing new phellinus igniarius anticancer drugs in a series, and the demand for phellinus igniarius is increasing. So that the wild phellinus igniarius resources are extremely rare and almost extinct in predation collection of medicine farmers in various domestic producing areas, and therefore, the artificial cultivation for making up the deficiency of the wild resources is very important.
Research on high-yield cultivation technology of phellinus linteus starts early in japan and korea, and a mature technology has been formed. Since the research of phellinus linteus in China arose since the end of the last century, because high-end cultivation techniques of Japan and Korea are used as patents, the protection is strict, and at present, the related research strength in China is weak, the artificial cultivation technique is not mature enough, and certain difficulties exist in the aspects of phellinus linteus strain breeding, culture medium screening, hypha culture and fruiting body growth and culture management. Although a few types of phellinus linteus have been successfully cultivated, the phellinus linteus is difficult to cultivate manually, has long growth period, is easy to be polluted by mixed bacteria, cannot realize standardized, industrialized and intelligent cultivation, and has poor quality, so that the phellinus linteus cannot meet the market demand. The intelligent cultivation of phellinus igniarius is still blank.
For example, patent publication No. CN108264390A discloses a method for industrially cultivating phellinus linteus fruiting bodies, which requires inoculation for 6 months to pick up the fruiting bodies, has a long cultivation period, and requires relatively expensive grain seeds as the main raw material of the cultivation medium; in addition, the grain seeds are adopted as culture medium, and the grain seeds can grow and develop, so that the nutrition loss is very large, and the accumulation of active ingredients is not facilitated.
The invention discloses a phellinus igniarius cultivation method which mainly utilizes pueraria lobata residues as an auxiliary material to be added into a basswood fungus bag, so that phellinus igniarius hyphae can be planted and germinate rapidly, the pollution rate of the fungus bag is reduced, and the yield of phellinus igniarius fungus bags is improved. However, the method does not solve the problems that the growth process of phellinus igniarius sporocarp is slow and the fruiting uniformity is not high.
The invention discloses an artificial high-yield cultivation method of phellinus igniarius and application of phellinus igniarius sporocarp, which are applied to the invention with the application number of 200410041704. X. The addition of auxin in the culture medium does not fundamentally solve the problem of the culture medium, and the addition of auxin has influence on the quality of phellinus igniarius and is not beneficial to the natural growth of phellinus igniarius.
The invention patent with application number 201710265348.7 discloses a cultivation method for improving the content of phellinus igniarius polysaccharide, which comprises the steps of matrix material preparation, bagging, high-temperature sterilization, inoculation, hypha culture, field fruiting management and picking; the increase in polysaccharide content by 10-15% is not satisfactory.
The invention patent of publication No. CN107996288A discloses a method for under-forest wild-like ecological cultivation of Phellinus linteus, which secretly simulates the wild growth environment of Phellinus linteus to obtain high-quality Phellinus linteus with considerable yield; provides a way for the wild-simulated ecological cultivation of phellinus igniarius under the forest. However, the content of flavone in phellinus igniarius cultivated by the technical scheme disclosed by the invention is only about 2.7%, and the obtained phellinus igniarius is low in quality.
In addition, about 1000-2000 million tons of plant organic matters are formed every year all over the world, wherein half of the organic matters are cellulose substances, and about 7 million tons of crop residues are formed only in agricultural production every year in China, so that the wastes are not fully utilized and pollute the environment.
Therefore, it is a technical problem to be urgently needed to develop a high-efficiency cultivation technique of phellinus igniarius, which has a short cultivation period, can be produced in a standardized, industrial and intelligent manner, effectively solves the pollution problem in the cultivation process, keeps the product quality stable, and greatly reduces the production cost.
Disclosure of Invention
In view of the above, the invention provides a method for performing real-time monitoring and intelligent control on a culture laboratory through a mobile phone terminal, so as to achieve accurate control on a culture environment, thereby realizing standardized, industrialized and intelligent production of cultivation management phellinus linteus.
In order to achieve the purpose, the invention adopts the following technical scheme: a method for improving the cultivation efficiency of phellinus igniarius through intelligent cultivation management in a laboratory comprises the following steps:
(1) quality improvement and optimization of a solid plate culture medium of a phellinus igniarius mother strain: inoculating the activated phellinus igniarius slant mother culture to a phellinus igniarius mother culture solid plate culture medium, culturing at the temperature of 26 ℃ for 7-10 days for quality improvement and optimization, and selecting strains which are high in growth speed, dense and vigorous and have regular edges for later use;
(2) culturing a phellinus igniarius liquid strain propagation culture medium: inoculating strains subjected to quality improvement optimization of the solid plate culture medium into a phellinus igniarius liquid strain propagation culture medium under an aseptic condition, and performing intermittent shaking culture to obtain high-quality and stable liquid phellinus igniarius strains;
(3) the mulberry twig bag material optimizing culture medium: inoculating the uniformly shaken liquid strain by using a polyethylene bag, and then transferring the liquid strain into a spawn running chamber to culture under the conditions of constant temperature, constant humidity and constant carbon dioxide;
(4) spawn running management: sterilizing a inoculation chamber, a spawn running chamber and a fruiting chamber periodically before inoculation;
ventilating for 10min for 2 times every day after 30-40% of fungus bag is inoculated;
inoculating and culturing for 30-35d, when the hypha in the culture bag has large amount of protruded yellowish tumor-shaped protrusions, transferring the bag to a fruiting chamber with constant temperature of 26-28 deg.C, constant humidity of 90-95%, and constant CO2Culturing for 10 days under the condition of 500-600ppm concentration and then performing opening fruiting management;
(5) and (3) fruiting management: opening an opening at the middle lower part of the fungus bag, performing intelligent management in a mobile phone remote control laboratory, and performing accurate control on cultivation conditions, wherein the temperature is controlled at 26-28 ℃, the humidity is controlled at 90-95%, and CO is added2The concentration is 600-;
ventilating for 3 times every day, each time for 20-30min, illuminating for 8-10 hr every day, and culturing for 35-40d to obtain fresh Phellinus Linteus fruiting body when the Phellinus Linteus fruiting body turns from orange to deep yellow and the edge is hardened.
Further, the solid plate culture medium for each 1L of Phellinus linteus mother strain comprises: 200-230g of potato, 20-24g of glucose, 0.2-0.4g of lignin and 1.1-1.3g of MgSO4，1.4-1.6gKH2PO41.0-3.0ml of wood vinegar, 16.0-18.0g of agar and the balance of water; and has a pH of 6.0。
Adopt above-mentioned further beneficial effect to lie in: according to the invention, the quality of the strain can be effectively improved and optimized by additionally adding lignin and pyroligneous into the solid plate culture medium of the phellinus igniarius mother strain.
Further, the culture medium for the expansion and propagation of each 1L of phellinus linteus liquid strain comprises: 200-230g of potato, 18-20g of water-soluble soybean cake powder, 0.03-0.05g of melatonin and 1.1-1.3g of MgSO4，1.4-1.6gKH2PO40.4-0.6ml of wood vinegar and the balance of water; and the pH was 7.0.
Adopt above-mentioned further beneficial effect to lie in: in the invention, melatonin and wood vinegar are additionally added into the phellinus igniarius liquid strain propagation culture medium, so that the obtained liquid phellinus igniarius strain is more excellent and stable.
Further, the mulberry twig bag material optimized culture medium comprises the following raw materials in percentage by mass: 40-50% of mulberry twig sawdust, 12-15% of bagasse, 12-15% of pleurotus eryngii fungus residues, 15-20% of wheat bran, 7-10% of cordyceps militaris cake residues, 1.0% of gypsum powder and 1.0% of lime powder.
Adopt above-mentioned further beneficial effect to lie in: bagasse, pleurotus eryngii fungus residues and cordyceps militaris cake residues are added into the mulberry branch bag material optimized culture medium, so that nutrition can be further provided for phellinus igniarius sporocarp, and biological activity accumulation is promoted, so that the quality of phellinus igniarius is improved.
Further, the shake culture in the step (2) is: and (3) performing static culture for 1d at the temperature of 26 ℃ in a dark environment, and performing high-speed oscillation for 22-23 h at the speed of 160-180 r/min and low-speed oscillation for 1-2 h at the speed of 60-80 r/min every day from day 2 for 6-8 d.
Further, the polyethylene bag in the step (3) is a polyethylene fungus bag of 17CM × 33CM × 0.05CM, a puncher is inserted into the middle of the polyethylene bag, the top end wall of the puncher is 13CM and is provided with a hole, and a fungus ring and a fungus cover are sleeved on the top end wall of the puncher for sealing;
the polyethylene bag is sterilized at 121 deg.C under high pressure for 3.0-3.5h before inoculation, and the puncher is taken out after cooling to room temperature.
Further, the inoculation amount of each polyethylene bag in the step (3) is 15-20 ml;
the constant temperature, constant humidity and constant oxygenThe carbonization conditions comprise 25-27 deg.C, 60-65% humidity and CO2The concentration is 100-150 ppm.
Further, the sterilization method in the step (4) is to perform ultraviolet lamp irradiation for 22 hours and ozone sterilization for 2 hours continuously for 25 days.
Further, in the step (5), the mobile phone remotely controls the intelligent management of the laboratory, and the cultivation conditions are remotely, flexibly and accurately controlled all weather, wherein the conditions of constant temperature, constant humidity and constant carbon dioxide are that the temperature is 26-28 ℃, the humidity is 90-95%, and CO is used for controlling the temperature, the humidity and the carbon dioxide content2The concentration is 600-; ventilating for 3 times every day, 20-30min each time, and illuminating for 8-10h each day.
The opening in the step (5) is a horseshoe-shaped opening with the length of 8.0-10.cm and the width of 2.5-3.0 cm.
Still further, the above method further comprises step (6): and (4) continuously ventilating and heating the cultivation room, reducing the humidity of the cultivation room to below 50%, air-drying the surface of the sporocarp for 2 days, and uniformly harvesting to obtain the mature phellinus igniarius sporocarp.
The invention has the beneficial effects that: in the method, the cultivation time of the phellinus igniarius sporocarp is 110 days from inoculation, the forming rate of the sporocarp can reach more than 94 percent, the production speed of the sporocarp is high, 70 days are spent from cutting to harvesting of fungus bags, the yield per unit (dry) is 30-40g, the weight of an average single-package dry product can reach 32g, and the sporocarp looks: smooth surface, uniform appearance and good commodity. The active ingredients of the intelligent cultivated phellinus igniarius sporocarp in a laboratory are obviously improved compared with the contents of polysaccharide, ergosterol and triterpene in the artificially cultivated phellinus igniarius sporocarp in a factory and the wild phellinus igniarius sporocarp.
The method has the advantages of short culture period of phellinus igniarius mycelium and sporocarp, high biotransformation rate, high yield, excellent quality and low pollution rate, and can realize annual standardized and large-scale production. The solid plate quality improvement optimization culture medium of the phellinus igniarius mother culture enables phellinus igniarius hyphae to grow densely and vigorously and quickly; the liquid strain propagation culture medium ensures that phellinus igniarius hypha balls are uniform, the propagation time is short, and the growth vigor is vigorous; the mulberry branch bag material optimized culture medium has rich nutrition of active ingredients, and the bag filling time of hyphae is shortened to about 40 d; the culture period of the phellinus igniarius sporocarp after the fungus bag is opened is shortened to about 70 days, the biotransformation rate is high, the sporocarp yield is high, and the contents of polysaccharide, ergosterol, triterpene and the like are obviously improved.
Drawings
FIG. 1 is a schematic diagram of the structure of the laboratory intelligent cultivation management bacteria room and the cultivation room.
FIG. 2 is a morphological diagram of Phellinus linteus fruiting body before harvesting in the example.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The structure schematic diagram of the laboratory intelligent cultivation management bacteria room and the cultivation room is shown in figure 1,
intelligent cultivation facilities:
a. the intelligent cultivation house:
the requirements of the cultivation room are as follows: the height is 6.5.0-7.0m, the length is 15-20m, the width is 12-15m, the cultivation room comprises an inoculation room, a spawn running room and a fruiting room, the inoculation room, the spawn running room and the fruiting room are connected through inner partition doors, the spawn running room and the fruiting room are connected side by side, the periphery and the top of the room are in a steel frame structure and are covered by a 7cm rear insulation board, and the ground is treated by resin lacquer glue;
the cultivation room is internally provided with ventilation equipment, temperature and humidity control equipment, illumination equipment, ultraviolet ozone sterilization equipment, intelligent main control equipment, a wired network, a wireless network, monitoring equipment and a double-sided superclean bench, and all-weather remote control and adjustment can be carried out on each equipment of the cultivation room through software corresponding to the intelligent main control equipment for 24 hours, so that the cultivation management conditions can be most closely and accurately fitted with the growth habitat of wild phellinus igniarius, and the advantages of shortening the growth period and improving the quality of phellinus igniarius are achieved;
b. a spawn running bed and a cultivation bed: the cultivation bed frame is built by stainless steel to fungus bed and cultivation bed, and the movable pulley is installed at four corners, is convenient for conveniently adjust and put. The bed width is 1.0-1.2m, the length is 1.5-1.6m, the number of bed frame layers is 5-6, each layer adopts a 6cm x 6cm fine grid tray, the bottom layer of the bed frame is 0.3-0.4m away from the ground, the distance between two adjacent layers is 0.3-0.4m, and a passageway with the length of 1.2-1.5m is reserved between two adjacent bed frames;
example 2
Quality improvement and optimization of a solid plate culture medium of a phellinus igniarius mother strain: inoculating the activated phellinus igniarius slant mother culture to a phellinus igniarius mother culture solid plate culture medium under aseptic conditions, culturing at 26 ℃ for 10 days for quality improvement and optimization, and selecting solid phellinus igniarius strains which are high in growth speed, dense and vigorous and neat in edges for later use.
The solid plate culture medium comprises the following components: 230g/L potato, 23g/L glucose, 0.3g/L lignin, 1.2g/LMgSO4，1.5g/LKH2PO41.5ml/L of wood vinegar, 18.0g/L of agar and the balance of water, and the pH is adjusted to 6.0.
And (3) performing discontinuous propagation culture on phellinus igniarius liquid strains: the liquid propagation culture medium is sterilized under high pressure for 30min at the temperature of 121 ℃, and is cooled in an aseptic environment. Inoculating 10 Phellinus Linteus strains with diameter of 0.6cm into liquid culture medium under aseptic condition, performing intermittent shake culture, standing at 26 deg.C under aseptic dark condition for 1d, starting shake culture on day 2, performing high speed shake at 180r/min for 23h and 60r/min for 1h every day, and performing shake culture for 7d to obtain high-quality and stable Phellinus Linteus strains.
The liquid propagation culture medium comprises 230g/L of potato, 18g/L of water-soluble soybean cake powder, 0.03g/L of melatonin and 1.1g/LMgSO4，1.4g/LKH2PO40.5ml/L of wood vinegar and the balance of water, and the pH is adjusted to 7.0.
The mulberry twig bag material optimizing culture medium: bagging with 17CM × 33CM × 0.05CM polyethylene fungus bag, inserting a plastic puncher with holes on the top end wall of 13CM long in the middle after bagging, sleeving a fungus ring and a fungus cover for sealing; the temperature is 121 ℃, the autoclaving time is 3.0 hours, the puncher is taken out of a bidirectional superclean workbench of an inoculation room after the temperature is cooled to the room temperature, 15ml of uniformly shaken liquid strains are inoculated by an inoculation gun, a strain cover is quickly covered, and the strain is transferred into a sterile spawn running room to be cultured under the conditions of constant temperature, constant humidity and constant carbon dioxide.
The mulberry twig bag material optimized culture medium comprises 45% of mulberry twig sawdust, 15% of bagasse, 12% of pleurotus eryngii fungus residues, 18% of wheat bran, 8% of cordyceps militaris cake residues, 1.0% of gypsum powder and 1.0% of lime powder, and the water content of the phellinus igniarius culture medium is 55%.
The intelligent spawn running of cell-phone, the management is cultivateed to the fruiting: 25d before the inoculation, inoculation room, spawn running room, fruiting room, through the first hat cell-phone APP networking control of smart mobile phone application intelligence, set for every day and start 22 hours ultraviolet lamp and shine and 2 hours ozone disinfection, 25d in succession carry out the disinfection of thoroughly disinfecting of alternation to the cultivation room. The inoculated fungus bags are intelligently and accurately controlled at the temperature of 25 ℃, the humidity of 60 percent and CO2Culturing in a spawn running room with the concentration of 100 ppm. Ventilating the fungus growing room for 2 times every day after about one third of fungus growing in the fungus bag, wherein each time is 10min, and sufficient oxygen is provided to facilitate normal, rapid and vigorous growth of hyphae. Culturing for 35 days, when the hypha in the culture bag has a large amount of protruded yellowish nodular protrusions, transferring the bag to a fruiting room, keeping the temperature at 27 deg.C, keeping the humidity at 90%, and adapting to 500ppm in a constant carbon dioxide environment for 10 days, and performing fruiting management after opening. Opening 1U-shaped opening with length of 8.0-10 cm and width of 2.5-3.0cm at the middle lower part of the fungus bag with scalpel, leaving 8cm space between the fungus bag and the fungus bag, controlling temperature at 26 deg.C, humidity at 93%, CO2 concentration at 800ppm, culturing for 18 days, maintaining internal illumination intensity at 90lux, and illuminating for 4hr per day. Ventilating for 3 times per day after 12 days, 30min each time, and lighting for 8hr each day. And continuing culturing for 40 days, and harvesting to obtain fresh Phellinus linteus fruiting body when the Phellinus linteus fruiting body turns from orange to deep yellow and the edge is hardened to show that the Phellinus linteus fruiting body is mature. At this time, after the humidity of the cultivation room is reduced by ventilating and increasing the temperature of the cultivation room, the humidity is reduced to be below 50%, the surface of the sporocarp is air-dried for 2 days, and then the sporocarp is collected uniformly.
In the whole process, the cultivation time of phellinus igniarius sporocarp is 117 days from inoculation, the forming rate of the sporocarp can reach more than 95%, the production speed of the sporocarp is high, 72 days are spent from cutting to harvesting of fungus bags, the yield per unit (dry) is 26-42 g, the weight of an average single-package dry product can reach 30g, and the sporocarp looks: smooth surface, uniform appearance and good commodity. The method comprises the following steps of intelligently cultivating active ingredients of phellinus igniarius sporocarp in a laboratory: in the embodiment, the contents of polysaccharide, ergosterol and total triterpene in the active ingredients of the cultivated phellinus igniarius sporocarp are 8.237g/100g, 2.756mg/100g and 2.337g/100g respectively.
Example 3
Quality improvement and optimization of a solid plate culture medium of a phellinus igniarius mother strain: inoculating the activated phellinus igniarius slant mother culture to a phellinus igniarius mother culture solid plate culture medium under aseptic conditions, culturing at 26 ℃ for 10 days for quality improvement and optimization, and selecting solid phellinus igniarius strains which are high in growth speed, dense and vigorous and neat in edges for later use.
The solid plate culture medium comprises the following components: 200g/L potato, 24g/L glucose, 0.2g/L lignin, 1.1g/LMgSO4，1.4g/LKH2PO42.0ml/L of wood vinegar, 17.0g/L of agar and the balance of water, and the pH is adjusted to 6.0.
And (3) performing discontinuous propagation culture on phellinus igniarius liquid strains: the liquid propagation culture medium is sterilized under high pressure for 30min at the temperature of 121 ℃, and is cooled in an aseptic environment. Inoculating 10 Phellinus Linteus strains with diameter of 0.6cm into liquid culture medium under aseptic condition, performing intermittent shake culture, standing at 26 deg.C under aseptic dark condition for 1d, starting shake culture on day 2, shaking at high speed of 170r/min for 23h and low speed of 80r/min for 1h every day, and shake culture for 8 days to obtain high-quality and stable Phellinus Linteus strains.
The liquid propagation culture medium comprises 220g/L potato, 20g/L water-soluble soybean cake powder, 0.04g/L melatonin, and 1.2g/LMgSO4，1.5g/LKH2PO40.6ml/L of wood vinegar and the balance of water, and the pH is adjusted to 7.0.
The mulberry twig bag material optimizing culture medium: bagging with 17CM × 33CM × 0.05CM polyethylene fungus bag, inserting a plastic puncher with holes on the top end wall of 13CM long in the middle after bagging, sleeving a fungus ring and a fungus cover for sealing; the temperature is 121 ℃, the autoclaving time is 3.0 hours, the puncher is taken out of a bidirectional superclean workbench of an inoculation room after the temperature is cooled to the room temperature, 18ml of uniformly shaken liquid strains are inoculated by an inoculation gun, a strain cover is quickly covered, and the strain is transferred into a sterile spawn running room to be cultured under the conditions of constant temperature, constant humidity and constant carbon dioxide.
The mulberry branch bag material optimized culture medium comprises the components of 50% of mulberry branch sawdust, 12% of bagasse, 13% of pleurotus eryngii fungus residues, 15% of wheat bran, 8% of cordyceps militaris cake residues, 1.0% of gypsum powder and 1.0% of lime powder, and the water content of the phellinus igniarius culture medium is 55%.
The intelligent spawn running of cell-phone, the management is cultivateed to the fruiting: 25 days before inoculation, an inoculation chamber, a spawn running chamber and a fruiting chamber are controlled through a smart phone network, 22-hour ultraviolet lamp irradiation and 2-hour ozone disinfection are set and started every day, 25 days are continuously carried out, and the cultivation chamber is alternately and thoroughly sterilized and disinfected. The inoculated fungus bags are intelligently and accurately controlled at the temperature of 28 ℃, the humidity of 65 percent and CO2Culturing in a spawn running room with the concentration of 150 ppm. The fungus bag is ventilated twice in the fungus growing room every day after about one third of the fungus growing time, and each time is 10min, so that sufficient oxygen is provided to facilitate normal, rapid and vigorous growth of hyphae. Culturing for 30 days, when the hyphae in the culture bag have a large amount of protruded yellowish nodular protrusions, transferring the bag to a fruiting room, keeping the temperature at 26 ℃, keeping the humidity at 92% and adapting to 600ppm in a constant carbon dioxide environment for 10 days, and then performing opening fruiting management. Opening 1U-shaped opening with length of 8.0-10 cm and width of 2.5-3.0cm at the middle and lower part of the fungus bag with scalpel, leaving 10cm space between the fungus bag and the fungus bag, controlling temperature at 28 deg.C, humidity at 90%, and CO at 90%2The concentration is 1200ppm, and after culturing for 15d, the internal illumination intensity is maintained at 80lux, and the illumination time is 3hr per day. Ventilating for 3 times a day after culturing for 15d, each time for 30min, and lighting for 9h each day. And continuing to culture for 38 days, and harvesting to obtain fresh Phellinus linteus fruiting body when the Phellinus linteus fruiting body turns from orange to deep yellow and the edge is hardened to show that the Phellinus linteus fruiting body is mature. At this time, after the humidity of the cultivation room is reduced by ventilating and increasing the temperature of the cultivation room, the humidity is reduced to be below 50%, the surface of the sporocarp is air-dried for 2 days, and then the sporocarp is collected uniformly.
In the whole process, the cultivation time of phellinus igniarius sporocarp is 110d from inoculation, the forming rate of the sporocarp can reach more than 94%, the production speed of the sporocarp is high, the total time from cutting to harvesting of fungus bags is 70d, the yield per unit (dry) is 30-40g, the weight of an average single-package dry product can reach 32g, and the sporocarp looks: smooth surface, uniform appearance and good commodity.
The method comprises the following steps of intelligently cultivating active ingredients of phellinus igniarius sporocarp in a laboratory: the contents of polysaccharide, ergosterol and total triterpene in the active ingredients of the sporocarp of the phellinus igniarius cultivated intelligently in the embodiment are 7.954g/100g, 22.645mg/100g and 2.280g/100g respectively.
Example 4
Quality improvement and optimization of a solid plate culture medium of a phellinus igniarius mother strain: inoculating the activated phellinus igniarius slant mother culture to a phellinus igniarius mother culture solid plate culture medium under the aseptic condition, culturing for 10 days at the temperature of 25 ℃, upgrading and optimizing, and selecting solid phellinus igniarius strains which are high in growth speed, dense and vigorous and neat in edges for later use.
The solid plate culture medium comprises the following components: 220g/L potato, 20g/L glucose, 0.4g/L lignin, 1.3g/LMgSO4，1.6g/LKH2PO43.0ml/L of wood vinegar, 16.0g/L of agar and the balance of water, and the pH is adjusted to 6.0.
And (3) performing discontinuous propagation culture on phellinus igniarius liquid strains: the liquid propagation culture medium is sterilized under high pressure for 30min at the temperature of 121 ℃, and is cooled in an aseptic environment. Inoculating 10 Phellinus Linteus strains with diameter of 0.6cm into liquid culture medium under aseptic condition, performing intermittent shake culture, standing at 26 deg.C under aseptic dark condition for 1d, starting shake culture on day 2, shaking at high speed of 160r/min for 22h and low speed of 75r/min for 2h every day, and shake culturing for 6d to obtain high-quality and stable Phellinus Linteus strains.
The liquid propagation culture medium comprises 200g/L potato, 19g/L water-soluble soybean cake powder, 0.05g/L melatonin, and 1.3g/LMgSO4，1.6g/LKH2PO40.4ml/L of wood vinegar and the balance of water, and the pH is adjusted to 7.0.
The mulberry twig bag material optimizing culture medium: bagging with 17CM × 33CM × 0.05CM polyethylene fungus bag, inserting a plastic puncher with holes on the top end wall of 13CM long in the middle after bagging, sleeving a fungus ring and a fungus cover for sealing; the temperature is 121 ℃, the autoclaving time is 3.0 hours, the puncher is taken out of a bidirectional superclean workbench of an inoculation room after the temperature is cooled to room temperature, 20ml of uniformly shaken liquid strains are inoculated by an inoculation gun, a strain cover is quickly covered, and the strain is transferred into a sterile spawn running room to be cultured under the conditions of constant temperature, constant humidity and constant carbon dioxide.
The mulberry twig bag material optimized culture medium comprises 40% of mulberry twig sawdust, 14% of bagasse, 14% of pleurotus eryngii fungus residues, 20% of wheat bran, 10% of cordyceps militaris cake residues, 1.0% of gypsum powder and 1.0% of lime powder, and the water content of the phellinus igniarius culture medium is 55%.
The intelligent spawn running of cell-phone, the management is cultivateed to the fruiting: 25d before the inoculation, inoculation room, spawn running room, fruiting room, through the first hat cell-phone APP networking control of smart mobile phone application intelligence, set for every day and start 22 hours ultraviolet lamp and shine and 2 hours ozone disinfection, 25d in succession carry out the disinfection of thoroughly disinfecting of alternation to the cultivation room. The inoculated fungus bags are intelligently and accurately controlled at the temperature of 26 ℃, the humidity of 63 percent and CO2Culturing under the environment condition of a spawn running room with the concentration of 130 ppm. The fungus bag is ventilated twice in the fungus growing room every day after about one third of the fungus growing time, and each time is 10min, so that sufficient oxygen is provided to facilitate normal, rapid and vigorous growth of hyphae. Culturing for 33 days, when the hyphae in the cultivation bag have a large amount of protruded faint yellow tumor-shaped protrusions, moving the fungus bag to a fruiting room, keeping the temperature at 28 ℃, keeping the humidity at 95%, adapting to 550ppm in a constant carbon dioxide environment for 10 days, and then carrying out opening fruiting management. Opening 1U-shaped opening with length of 8.0-10 cm and width of 2.5-3.0cm at the middle and lower part of the fungus bag with scalpel, leaving 12.0cm space between the fungus bag and the fungus bag, controlling temperature at 27 deg.C, humidity at 95%, CO2 concentration at 600ppm, culturing for 20 days, maintaining internal illumination intensity at 150lux, and illuminating for 5h per day. Ventilating for 3 times per day after culturing for 10 days, each time for 30min, and lighting for 10hr per day. And continuing culturing for 35d, and harvesting to obtain fresh Phellinus linteus fruiting body when the Phellinus linteus fruiting body turns from orange to deep yellow and the edge is hardened to show that the Phellinus linteus fruiting body is mature. At this time, after the humidity of the cultivation room is reduced by ventilating and increasing the temperature of the cultivation room, the humidity is reduced to be below 50%, the surface of the sporocarp is air-dried for 2 days, and then the sporocarp is collected uniformly.
In the whole process, the cultivation time of phellinus igniarius sporocarp is 110d from inoculation, the forming rate of the sporocarp can reach more than 94%, the production speed of the sporocarp is high, 67d is totally obtained from cutting to harvesting of fungus bags, the yield per unit (dry) is 30-40g, the weight of an average single-package dry product can reach 32g, and the sporocarp looks: smooth surface, uniform appearance and good commodity.
The method comprises the following steps of intelligently cultivating active ingredients of phellinus igniarius sporocarp in a laboratory: the content of polysaccharide, ergosterol and total triterpene in the active ingredients of the sporocarp of phellinus igniarius cultivated intelligently in the embodiment is 8.067g/100g, 2.733mg/100g and 2.316g/100g respectively.
Test example 1
The results of testing examples 2-4 and comparison of the contents of active ingredients in different management modes purchased from the market are shown in table 1.
Table 1 comparison table of active ingredient contents for different management modes
Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made to the above embodiments by those of ordinary skill in the art within the scope of the present invention.
Claims (10)
1. A method for improving the cultivation efficiency of phellinus igniarius through intelligent cultivation management in a laboratory is characterized by comprising the following steps:
(1) quality improvement and optimization of a solid plate culture medium of a phellinus igniarius mother strain: inoculating the activated phellinus igniarius slant mother culture to a phellinus igniarius mother culture solid plate culture medium, culturing at the temperature of 26 ℃ for 7-10 days for quality improvement and optimization, and selecting strains which are high in growth speed, dense and vigorous and have regular edges for later use;
(2) culturing a phellinus igniarius liquid strain propagation culture medium: inoculating strains subjected to quality improvement optimization of the solid plate culture medium into a phellinus igniarius liquid strain propagation culture medium under an aseptic condition, and performing intermittent shaking culture to obtain high-quality and stable liquid phellinus igniarius strains;
(3) the mulberry twig bag material optimizing culture medium: inoculating the uniformly shaken liquid strain by using a polyethylene bag, and then transferring the liquid strain into a spawn running chamber to culture under the conditions of constant temperature, constant humidity and constant carbon dioxide;
(4) spawn running management: sterilizing a inoculation chamber, a spawn running chamber and a fruiting chamber periodically before inoculation;
ventilating for 10min for 2 times every day after 30-40% of fungus bag is inoculated;
inoculating and culturing for 30-35d until the mycelium in the culture bag appears a large amount of protruded yellowish tumorWhen the mushroom bag protrudes, the mushroom bag is moved to a mushroom growing chamber, the temperature of the mushroom growing chamber is kept at 26-28 ℃, the humidity is kept at 90-95%, and CO is kept constant2Culturing for 10 days under the condition of 500-600ppm concentration and then performing opening fruiting management;
(5) and (3) fruiting management: opening an opening at the middle lower part of the fungus bag, performing intelligent management in a mobile phone remote control laboratory, and performing accurate control on cultivation conditions, wherein the temperature is 26-28 ℃, the humidity is 90-95%, and CO is used as a carrier2The concentration is 600-; ventilating for 3 times every day, each time for 20-30min, illuminating for 8-10 hr every day, and culturing for 35-40d to obtain fresh Phellinus Linteus fruiting body when the Phellinus Linteus fruiting body turns from orange to deep yellow and the edge is hardened.
2. The method for improving the cultivation efficiency of phellinus igniarius through intelligent laboratory cultivation management according to claim 1, wherein each 1L of phellinus igniarius mother culture solid plate culture medium comprises: 200-230g of potato, 20-24g of glucose, 0.2-0.4g of lignin and 1.1-1.3g of MgSO4，1.4-1.6gKH2PO41.0-3.0ml of wood vinegar, 16.0-18.0g of agar and the balance of water; and a pH of 6.0.
3. The method for improving the cultivation efficiency of phellinus igniarius through the intelligent laboratory cultivation management according to claim 1, wherein the expanding culture medium for every 1L of phellinus igniarius liquid spawn comprises: 200-230g of potato, 18-20g of water-soluble soybean cake powder, 0.03-0.05g of melatonin and 1.1-1.3g of MgSO4，1.4-1.6gKH2PO40.4-0.6ml of wood vinegar and the balance of water; and the pH was 7.0.
4. The method for improving the cultivation efficiency of phellinus igniarius through the intelligent laboratory cultivation management according to claim 1, wherein the mulberry twig bag material optimizing culture medium comprises the following raw materials in percentage by mass: 40-50% of mulberry twig sawdust, 12-15% of bagasse, 12-15% of pleurotus eryngii fungus residues, 15-20% of wheat bran, 7-10% of cordyceps militaris cake residues, 1.0% of gypsum powder and 1.0% of lime powder.
5. The method for improving the cultivation efficiency of phellinus igniarius through the intelligent laboratory cultivation management according to claim 1, wherein the shaking cultivation in the step (2) is as follows: and (3) performing static culture for 1d at the temperature of 26 ℃ in a dark environment, and performing high-speed oscillation for 22-23 h at the speed of 160-180 r/min and low-speed oscillation for 1-2 h at the speed of 60-80 r/min every day from day 2 for 6-8 d.
6. The method for improving the cultivation efficiency of phellinus igniarius through intelligent laboratory cultivation management according to claim 1, wherein in the step (3), the polyethylene bag is a 17CM x 33CM x 0.05CM polyethylene fungus bag, a hole puncher is inserted in the middle of the polyethylene bag, the hole puncher is a hole in the top end wall of 13CM, and a fungus ring and a fungus cover sealing are sleeved on the hole;
the polyethylene bag is sterilized at 121 deg.C under high pressure for 3.0-3.5h before inoculation, and the puncher is taken out after cooling to room temperature.
7. The method for improving the cultivation efficiency of phellinus igniarius through intelligent laboratory cultivation management according to claim 1, wherein the disinfection method in the step (4) is ultraviolet lamp irradiation for 22 hours and ozone disinfection for 2 hours every day for 25 days continuously.
8. The method for improving the cultivation efficiency of Phellinus linteus by intelligent laboratory cultivation management as claimed in claim 1, wherein in step (5), the intelligent laboratory management is remotely controlled by mobile phone to flexibly and accurately control cultivation conditions in all weather, and the conditions of constant temperature, humidity, carbon dioxide and CO are 26-28 deg.C, 90-95% humidity2The concentration is 600-; ventilating for 3 times every day, 20-30min each time, and illuminating for 8-10h each day.
9. The method for improving the cultivation efficiency of phellinus igniarius through intelligent laboratory cultivation management according to claim 1 or 8, wherein the opening in step (5) is a horseshoe-shaped opening with the length of 8.0-10.cm and the width of 2.5-3.0 cm.
10. The method for improving the cultivation efficiency of phellinus igniarius through intelligent laboratory cultivation management according to claim 1, further comprising the step (6): and (4) continuously ventilating and heating the cultivation room, reducing the humidity of the cultivation room to below 50%, air-drying the surface of the sporocarp for 2 days, and uniformly harvesting to obtain the mature phellinus igniarius sporocarp.
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